Compounds containing s-n-valeryl-n-{[2&#39;-(1h-tetrazole-5-yl)-biphenyl-4-yl]-methyl}-valine and (2r,4s)-5-biphenyl-4-yl-4-(3-carboxy-propionylamino)-2-methyl-pentanoic acid ethyl ester moieties and cations

ABSTRACT

A compound comprising of the angiotensin receptor antagonist (ARB) valsartan, the neutral endopeptidase inhibitor (NEPi) (2R,4S)-5-biphenyl-4-yl-4-(3-carboxy-propionylamino)-2-methylpentanoic acid ethyl ester and one or more monovalent cations such as Na +  useful for the treatment of hypertension and/or heart failure.

RELATED APPLICATIONS

This application is a divisional application of U.S. application Ser.No. 16/579,581, filed Sep. 23, 2019, which is a continuation applicationof U.S. application Ser. No. 16/006,252, filed on Jun. 12, 2018, whichis a continuation application of U.S. application Ser. No. 15/187,872,filed on Jun. 21, 2016, which is a divisional application of U.S.application Ser. No. 14/311,788, filed on Jun. 23, 2014, now U.S. Pat.No. 9,388,134, which is a divisional application of U.S. applicationSer. No. 11/722,360, filed on Jan. 15, 2008, now U.S. Pat. No.8,877,938, which is a national stage application, filed under 35 U.S.C.§ 371, of International Application No. PCT/US06/43710, filed on Nov. 8,2006, which claims the benefit of and priority to U.S. ProvisionalApplication Nos. 60/822,086, filed Aug. 11, 2006, 60/789,332, filed Apr.4, 2006, 60/735,541, filed on Nov. 10, 2005, and 60/735,093, filed onNov. 9, 2005, the entire contents of each of which are incorporatedherein by reference in their entireties.

FIELD OF THE INVENTION

The present invention is directed to dual-acting compounds andcombinations of angiotensin receptor blockers and neutral endopeptidaseinhibitors, in particular a dual acting molecule wherein the angiotensinreceptor blocker and neutral endopeptidase inhibitor are linked vianon-covalent bonding, or supramolecular complexes of angiotensinreceptor blockers and neutral endopeptidase inhibitors, also describedas linked pro-drugs, such as mixed salts or co-crystals, as well as topharmaceutical combinations containing such a dual-acting compound orcombination, methods of preparing such dual-acting compounds and methodsof treating a subject with such a dual-acting compound or combination.Specifically, the invention is directed to a dual acting compound orsupramolecular complex of two active agents having the same or differentmodes of action in one molecule.

BACKGROUND OF THE INVENTION

Angiotensin II is a hormone that causes blood vessels to constrict.This, in turn, can result in high blood pressure and strain on theheart. It is known that angiotensin II interacts with specific receptorson the surface of target cells. Two receptor subtypes for angiotensinII, namely AT1 and AT2, have been identified thus far. In recent times,great efforts have been made to identify substances that bind to the AT1receptor. Angiotensin receptor blockers (ARBs, angiotensin IIantagonists) are now known to prevent angiotensin II from binding to itsreceptors in the walls of blood vessels, thereby resulting in lowerblood pressure. Because of the inhibition of the AT1 receptor, suchantagonists can be used, therefore, as anti-hypertensives or for thetreatment of congestive heart failure, among other indications.

Neutral endopeptidase (EC 3.4.24.11; enkephalinase; atriopeptidase; NEP)is a zinc-containing metalloprotease that cleaves a variety of peptidesubstrates on the amino side of hydrophobic residues [see Pharmacol Rev,Vol. 45, p. 87 (1993)]. Substrates for this enzyme include, but are notlimited to, atrial natriuretic peptide (ANP, also known as ANF), brainnatriuretic peptide (BNP), met- and leu-enkephalin, bradykinin,neurokinin A, endothelin-1 and substance P. ANP is a potent vasorelaxantand natriuretic agent [see J Hypertens, Vol. 19, p. 1923 (2001)].Infusion of ANP in normal subjects resulted in a reproducible, markedenhancement of natriuresis and diuresis, including increases infractional excretion of sodium, urinary flow rate and glomerularfiltration rate [see J Clin Pharmacol, Vol. 27, p. 927 (1987)]. However,ANP has a short half-life in circulation, and NEP in kidney cortexmembranes has been shown to be the major enzyme responsible fordegrading this peptide [see Peptides, Vol. 9, p. 173 (1988)]. Thus,inhibitors of NEP (neutral endopeptidase inhibitors, NEPi) shouldincrease plasma levels of ANP and, hence, are expected to inducenatriuretic and diuretic effects.

While substances, such as angiotensin receptor blockers and neutralendopeptidase inhibitors may be useful in the control of hypertension,essential hypertension is a polygenic disease and is not alwayscontrolled adequately by monotherapy. Approximately 333 million adultsin economically developed countries and about 65 million Americans (1 in3 adults) had high blood pressure in 2000 [see Lancet, Vol. 365, p. 217(2005); and Hypertension, Vol. 44, p. 398 (2004)]. Prolonged anduncontrolled hypertensive vascular disease ultimately leads to a varietyof pathological changes in target organs, such as the heart and kidney.Sustained hypertension can lead as well to an increased occurrence ofstroke. Therefore, there is a strong need to evaluate the efficacy ofanti-hypertensive therapy, an examination of additional cardiovascularendpoints, beyond those of blood pressure lowering, to get furtherinsight into the benefits of combined treatment.

The nature of hypertensive vascular diseases is multifactorial. Undercertain circumstances, drugs with different mechanisms of action havebeen combined. However, just considering any combination of drugs havingdifferent modes of action does not necessarily lead to combinations withadvantageous effects. Accordingly, there is a need for efficaciouscombination therapy which does not have deleterious side effects.

SUMMARY OF THE INVENTION

In a first aspect, the present invention is directed to a dual-actingcompound, such as a supramolecular complex, comprising:

-   -   (a) an angiotensin receptor antagonist;    -   (b) a neutral endopeptidase inhibitor (NEPi); and optionally    -   (c) a pharmaceutically acceptable cation.

The present invention is also directed to a dual-acting compound, suchas a supramolecular complex, obtainable by:

-   -   (i) dissolving an angiotensin receptor antagonist and a neutral        endopeptidase inhibitor (NEPi) in a suitable solvent;    -   (ii) dissolving a basic compound of Cat in a suitable solvent,        wherein Cat is a cation;    -   (iii) combining the solutions obtained in steps (i) and (ii);    -   (iv) precipitation of the solid, and drying same to obtain the        dual-acting compound; or alternatively    -   obtaining the dual-acting compound by exchanging the solvent(s)        employed in steps (i) and (ii) by    -   (iva) evaporating the resulting solution to dryness;    -   (va) re-dissolving the solid in a suitable solvent; (via)        precipitation of the solid and drying same to obtain the        dual-acting compound.

The present invention is also directed to linked pro-drugs comprising:

(a) an angiotensin receptor antagonist or a pharmaceutically acceptablesalt thereof; and

(b) a NEPi or a pharmaceutically acceptable salt thereof, wherein theangiotensin receptor antagonist or a pharmaceutically acceptable saltthereof and the NEPi or a pharmaceutically acceptable salt thereof arelinked by a linking moiety.

The present invention is also directed to a combination comprising:

-   -   (a) a pharmaceutically acceptable salt of an angiotensin        receptor antagonist; and    -   (b) a pharmaceutically acceptable salt of a neutral        endopeptidase inhibitor (NEPi);

wherein the pharmaceutically acceptable salt of the angiotensin receptorantagonist and the NEPi is the same and is selected from a salt of Na, Kor NH₄.

In preferred embodiments, the angiotensin receptor antagonist and NEPihave acidic groups which facilitate formation of the dual actingcompound, such as the supramolecular complex of the present invention.

Preferably, the angiotensin receptor antagonist is selected from thegroup consisting of valsartan, losartan, irbesartan, telmisartan,eprosartan, candesartan, olmesartan, saprisartan, tasosartan, elisartanand combinations thereof.

In preferred embodiments, the NEPi is selected from the group consistingof: SQ 28,603;N—[N-[1(S)-carboxyl-3-phenylpropyl]-(S)-phenylalanyl]-(S)-isoserine;N—[N-[((1 S)-carboxy-2-phenyl)ethyl]-(S)-phenylalanyl]-β-alanine;N-[2(S)-mercaptomethyl-3-(2-methylphenyl)-propionyl]methionine;(cis-4-[[[1-[2-carboxy-3-(2-methoxyethoxy)propyl]-cyclopentyl]carbonyl]amino]-cyclohexanecarboxylicacid); thiorphan; retro-thiorphan; phosphoramidon; SQ 29072;N-(3-carboxy-1-oxopropyl)-(4S)-p-phenylphenylmethyl)-4-amino-2R-methylbutanoicacid ethyl ester;(S)-cis-4-[1-[2-(5-indanyloxycarbonyl)-3-(2-methoxyethoxy)propyl]-1-cyclopentanecarboxamido]-1-cyclohexanecarboxylicacid;3-(1-[6-endo-hydroxymethylbicyclo[2,2,1]heptane-2-exo-carbamoyl]cyclopentyl)-2-(2-methoxyethyl)propanoicacid;N-(1-(3-(N-t-butoxycarbonyl-(S)-prolylamino)-2(S)-t-butoxy-carbonylpropyl)cyclopentanecarbonyl)-O-benzyl-(S)-serinemethyl ester; 4-[[2-(mercaptomethyl)-1-oxo-3-phenylpropyl]amino]benzoicacid;3-[1-(cis-4-carboxycarbonyl-cis-3-butylcyclohexyl-r-1-carbamoyl)cyclopentyl]-2S-(2-methoxyethoxymethyl)propanoicacid; N-((2S)-2-(4-biphenylmethyl)-4-carboxy-5-phenoxyvaleryl)glycine;N-(1-(N-hydroxycarbamoylmethyl)-1-cyclopentanecarbonyl)-L-phenylalanine;(S)-(2-biphenyl-4-yl)-1-(1H-tetrazol-5-yl)ethylamino) methylphosphonicacid;(S)-5-(N-(2-(phosphonomethylamino)-3-(4-biphenyl)propionyl)-2-aminoethyl)tetrazole;β-alanine;3-[1,1′-biphenyl]-4-yl-N-[diphenoxyphosphinyl)methyl]-L-alanyl;N-(2-carboxy-4-thienyl)-3-mercapto-2-benzylpropanamide;2-(2-mercaptomethyl-3-phenylpropionamido)thiazol-4-ylcarboxylic acid;(L)-(1-((2,2-dimethyl-1,3-dioxolan-4-yl)-methoxy)carbonyl)-2-phenylethyl)-L-phenylalanyl)-β-alanine;N—[N-[(L)-[1-[(2,2-dimethyl-1,3-dioxolan-4-yl)-methoxy]carbonyl]-2-phenylethyl]-L-phenylalanyl]-(R)-alanine;N—[N-[(L)-1-carboxy-2-phenylethyl]-L-phenylalanyl]-(R)-alanine;N-[2-acetylthiomethyl-3-(2-methyl-phenyl)propionyl]-methionine ethylester; N-[2-mercaptomethyl-3-(2-methylphenyl)propionyl]-methionine;N-[2(S)-mercaptomethyl-3-(2-methylphenyl)propanoyl]-(S)-isoserine;N—(S)-[3-mercapto-2-(2-methylphenyl)propionyl]-(S)-2-methoxy-(R)-alanine;N-[1-[[1(S)-benzyloxycarbonyl-3-phenylpropyl]amino]cyclopentylcarbonyl]-(S)-isoserine;N-[1-[[1(S)-carbonyl-3-phenylpropyl]amino]-cyclopentylcarbonyl]-(S)-isoserine;1,1′-[dithiobis-[2(S)-(2-methylbenzyl)-1-oxo-3,1-propanediyl]]-bis-(S)-isoserine;1,1′-[dithiobis-[2(S)-(2-methylbenzyl)-1-oxo-3,1-propanediyl]]-bis-(S)-methionine;N-(3-phenyl-2-(mercaptomethyl)-propionyl)-(S)-4-(methylmercapto)methionine;N-[2-acetylthiomethyl-3-phenyl-propionyl]-3-aminobenzoic acid;N-[2-mercaptomethyl-3-phenyl-propionyl]-3-aminobenzoic acid;N-[1-(2-carboxy-4-phenylbutyl)-cyclopentane-carbonyl]-(S)-isoserine;N-[1-(acetylthiomethyl)cyclopentane-carbonyl]-(S)-methionine ethylester;3(S)-[2-(acetylthiomethyl)-3-phenyl-propionyl]amino-s-caprolactam;N-(2-acetylthiomethyl-3-(2-methylphenyl)propionyl)-methionine ethylester; and combinations thereof. Preferably, the dual-acting compound orcombination, in particular the supramolecular complex, is a mixed saltor a co-crystal. It is also preferred that the linked pro-drug is amixed salt or a co-crystal.

In a second aspect, the present invention is directed to pharmaceuticalcomposition comprising

-   -   (a) the aforementioned dual-acting compound or combination, such        as the aforementioned complex; and    -   (b) at least one pharmaceutically acceptable additive.

The present invention is also directed to pharmaceutical compositionscomprising a linked pro-drug comprising:

-   -   (a) an angiotensin receptor antagonist or a pharmaceutically        acceptable salt thereof;    -   (b) a NEPi or a pharmaceutically acceptable salt thereof,    -   wherein the angiotensin receptor antagonist or a        pharmaceutically acceptable salt thereof and the NEPi or a        pharmaceutically acceptable salt thereof are linked by a linking        moiety; and    -   (c) at least one pharmaceutically acceptable additive.

In a third aspect, the present invention is directed to a method ofpreparing a dual-acting compound, in particular a supramolecularcomplex, comprising

-   -   (a) an angiotensin receptor antagonist;    -   (b) a neutral endopeptidase inhibitor (NEPi); and optionally    -   (c) a pharmaceutically acceptable cation selected from the group        consisting of Na, K and NH₄;    -   said method comprising the steps of:    -   (i) dissolving an angiotensin receptor antagonist and a neutral        endopeptidase inhibitor (NEPi) in a suitable solvent;    -   (ii) dissolving a basic compound of Cat in a suitable solvent,        wherein Cat is a cation;    -   (iii) combining the solutions obtained in steps (i) and (ii);    -   (iv) precipitation of the solid, and drying same to obtain the        dual-acting compound; or alternatively obtaining the dual-acting        compound by exchanging the solvent(s) employed in steps (i)        and (ii) by    -   (iva) evaporating the resulting solution to dryness;    -   (va) re-dissolving the solid in a suitable solvent;    -   (via) precipitation of the solid and drying same to obtain the        dual-acting compound.

The present invention is also directed to a method of making a linkedpro-drug comprising:

-   -   (a) an angiotensin receptor antagonist or a pharmaceutically        acceptable salt thereof;    -   (b) a NEPi or a pharmaceutically acceptable salt thereof,        wherein the angiotensin receptor antagonist or a        pharmaceutically acceptable salt thereof and the NEPi or a        pharmaceutically acceptable salt thereof are linked by a linking        moiety; and comprising adding a linking moiety and a solvent to        a mixture of an angiotensin receptor antagonist and a NEPi; and    -   (d) isolating the linked pro-drug.

In a fourth aspect, this invention is directed to a method of treatingor preventing a disease or condition, such as hypertension, heartfailure (acute and chronic), congestive heart failure, left ventriculardysfunction and hypertrophic cardiomyopathy, diabetic cardiac myopathy,supraventricular and ventricular arrhythmias, atrial fibrillation,atrial flutter, detrimental vascular remodeling, myocardial infarctionand its sequelae, atherosclerosis, angina (unstable or stable), renalinsufficiency (diabetic and non-diabetic), heart failure, anginapectoris, diabetes, secondary aldosteronism, primary and secondarypulmonary hypertension, renal failure conditions, such as diabeticnephropathy, glomerulonephritis, scleroderma, glomerular sclerosis,proteinuria of primary renal disease, and also renal vascularhypertension, diabetic retinopathy, other vascular disorders, such asmigraine, peripheral vascular disease, Raynaud's disease, luminalhyperplasia, cognitive dysfunction (such as Alzheimer's), glaucoma andstroke comprising administering the afore-mentioned dual-acting compoundor combination, in particular the supramolecular complex, or theafore-mentioned linked pro-drug, preferably, the complex, to a subjectin need of such treatment.

BRIEF DESCRIPTION OF THE DRAWING

FIG. 1 shows a pictorial representation of the unit cell of thesupramolecular complex of trisodium[3-((1S,3R)-1-biphenyl-4-ylmethyl-3-ethoxycarbonyl-1-butylcarbamoyl)propionate-(S)-3′-methyl-2′-(pentanoyl{2″-(tetrazol-5-ylate)biphenyl-4′-ylmethyl}amino)butyrate]hemipentahydratecomprising two asymmetric units. The following color code is used:grey=carbon atom; blue=nitrogen atom; red=oxygen atom; violet=sodiumatom.

DETAILED DESCRIPTION

The present invention relates to a dual-acting compound or combination,in particular a supramolecular complex, or linked pro-drug or inparticular a supramolecular complex of two active agents with differentmechanisms of action, namely an angiotensin receptor antagonist and aneutral endopeptidase inhibitor, which can form a unique molecularentity for the treatment of patients with various cardiovascular and/orrenal diseases.

One embodiment of the invention is directed to a physical combinationcomprising:

-   -   (a) a pharmaceutically acceptable salt of an angiotensin        receptor antagonist; and    -   (b) a pharmaceutically acceptable salt of a neutral        endopeptidase inhibitor (NEPi); wherein the pharmaceutically        acceptable salt of the angiotensin receptor antagonist and the        NEPi is the same and is selected from a salt of Na, K or NH₄.

Specifically, it is preferred that the two active agents are combinedwith each other so as to form a single dual-acting compound, inparticular a supramolecular complex. By doing so, a new molecular orsupramolecular entity is formed having distinct properties different tothe above physical combination.

Thus, the present invention is directed to a dual-acting compound, inparticular a supramolecular complex, comprising:

-   -   (a) an angiotensin receptor antagonist;    -   (b) a neutral endopeptidase inhibitor (NEPi); and    -   (c) a pharmaceutically acceptable cation preferably selected        from the group consisting of Na, K and NH₄.

The present invention is also directed to a dual-acting compound, inparticular a supramolecular complex, obtainable by:

-   -   (i) dissolving an angiotensin receptor antagonist and a neutral        endopeptidase inhibitor (NEPi) in a suitable solvent;    -   (ii) dissolving a basic compound of Cat such as (Cat)OH,        (Cat)₂CO₃, (Cat)HCO₃ in a suitable solvent, wherein Cat is a        cation preferably selected from the group consisting of Na, K        and NH₄;    -   (iii) combining the solutions obtained in steps (i) and (ii);    -   (iv) precipitation of the solid, and drying same to obtain the        dual-acting compound; or alternatively    -   obtaining the dual-acting compound by exchanging the solvent(s)        employed in steps (i) and (ii) by    -   (iva) evaporating the resulting solution to dryness;    -   (va) re-dissolving the solid in a suitable solvent;    -   (via) precipitation of the solid and drying same to obtain the        dual-acting compound.

The present invention is further directed to linked pro-drugscomprising:

-   -   (a) an angiotensin receptor antagonist or a pharmaceutically        acceptable salt thereof; and    -   (b) a NEPi or a pharmaceutically acceptable salt thereof,    -   wherein the angiotensin receptor antagonist or a        pharmaceutically acceptable salt thereof and the NEPi or a        pharmaceutically acceptable salt thereof are linked by a linking        moiety.

The two components are each linked to a linking moiety thereby creatinga linked pro-drug.

Preferably, the linked pro-drug is substantially pure; as used herein,“substantially pure” refers to at least 90%, more preferably at least95% and most preferably at least 98% purity.

As one preferred embodiment of the present invention, the linkedpro-drug has a structure such that by linking the two components withthe linking moiety, a supramolecular complex is formed.

For the purpose of the present invention, the term “dual-actingcompound” is intended to describe that these compounds have twodifferent modes of action in one compound, one is the angiotensinreceptor blockade resulting from the ARB molecular moiety of thecompound and the other is the neutral endopeptidase inhibition resultingfrom the NEPi molecular moiety of the compound.

For the purpose of the present invention, the term “compound” isintended to describe a chemical substance comprising covalent bondswithin the two pharmaceutically active agents, the ARB and the NEPimolecular moieties, and non-covalent interactions between these twopharmaceutically active agents, the ARB and the NEPi molecular moieties.

Typically, hydrogen bonding can be observed between the twopharmaceutically active agents, the ARB and the NEPi molecular moieties.Ionic bonds can be present between the cation and one or both of the twopharmaceutically active agents, the ARB and the NEPi molecular moieties.Other types of bonds may also be present within the compound such as vander Waals forces. For illustrative purposes, the dual-acting compound ofthe present invention could be represented as follows:

(ARB)-(L)_(m)-(NEPi)

wherein L is a linking moiety, such as a cation or is a noncovalent bondand m is an integer from 1 or more. In other words the ARB and NEPimoiety can be connected via non-covalent bonds such as hydrogen bonding.Alternatively or additionally they may be connected via a linking moietysuch as a cation.

In one embodiment, the dual-acting compound may be considered to be alinked pro-drug, whereby the linking moiety, such as the cation, linkingthe two pharmaceutically active agents, the ARB and the NEPi, forms thepro-drug of these agents which are released once the linked pro-drug isingested and absorbed.

In a preferred embodiment, the dual-acting compound is a complex, inparticular a supramolecular complex.

For the purpose of the present invention, the term “supramolecularcomplex” is intended to describe an interaction between the twopharmaceutically active agents, the cations and any other entity presentsuch as a solvent, in particular water, by means of noncovalent,intermolecular bonding between them. This interaction leads to anassociation of the species present in the supramolecular complexdistinguishing this complex over a physical mixture of the species.

The noncovalent intermolecular bonding can be any interactions known inthe art to form such supramolecular complexes, such as hydrogen bonding,van der Waals forces and 71-71 stacking. Ionic bonds can also bepresent. Preferably, there exists ionic bonding and additionallyhydrogen bonding to form a network of interactions within the complex.The supramolecular complex exists preferably in the solid state but mayalso be present in liquid media. As a preferred embodiment of theinvention, the complex is crystalline and in this case is preferably amixed crystal or co-crystal.

Typically, the dual-acting compound, in particular the supramolecularcomplex shows properties such as melting point, IR spectrum etc. thatare different from a physical mixture of the species.

Preferably, the dual-acting compound, in particular the supramolecularcomplex, has a network of non-covalent bonds, in particular hydrogenbonds, between the two pharmaceutically active agents and any solvent,if present, preferably water. Moreover, it is preferred that thedual-acting compound, in particular the supramolecular complex, has anetwork of non-covalent bonds, in particular ionic and hydrogen bonds,between the two pharmaceutically active agents, the cation and anysolvent, if present, preferably water. The cation is preferablycoordinated to several oxygen ligands, thus, providing a linkage betweenthese oxygen ligands. The oxygen ligands come from the carbonyl andcarboxylate groups present in the two pharmaceutically active agents andpreferably also from any solvent, if present, preferably water.

The dual acting compound comprises a molecular moiety of an angiotensinreceptor antagonist. This means that a molecular moiety derived from anangiotensin receptor antagonist is participating in the build-up of thedual-acting compound. The angiotensin receptor antagonist is part of thecompound and connected to the NEP inhibitor directly or indirectly vianon-covalent bonds. For sake of convenience, throughout the application,the term “angiotensin receptor antagonist” will be used when describingthis part of the compound. Angiotensin receptor antagonists (ARBs)suitable for use in the present invention include, without limitation,valsartan, losartan, irbesartan, telmisartan, eprosartan, candesartan,olmesartan saprisartan, tasosartan, elisartan, the compound with thedesignation E-1477 of the following formula

the compound with the designation SC-52458 of the following formula

the compound with the designation the compound ZD-8731 of the followingformula

Suitable angiotensin II receptor antagonist also includes, but is notlimited to, saralasin acetate, candesartan cilexetil, CGP-63170,EMD-66397, KT3-671, LR-B/081, valsartan, A-81282, BIBR-363, BIBS-222,BMS-184698, candesartan, CV-11194, EXP-3174, KW-3433, L-161177,L-162154, LR-B/057, LY-235656, PD-150304, U-96849, U-97018, UP-275-22,WAY-126227, WK-1492.2K, YM-31472, losartan potassium, E-4177, EMD-73495,eprosartan, HN-65021, irbesartan, L-159282, ME-3221, SL-91.0102,Tasosartan, Telmisartan, UP-269-6, YM-358, CGP-49870, GA-0056, L-159689,L-162234, L-162441, L-163007, PD-123177, A-81988, BMS-180560,CGP-38560A, CGP-48369, DA-2079, DE-3489, DuP-167, EXP-063, EXP-6155,EXP-6803, EXP-7711, EXP-9270, FK-739, HR-720, ICI-D6888, ICI-D7155,ICI-D8731, isoteoline, KRI-1177, L-158809, L-158978, L-159874, LR B087,LY-285434, LY-302289, LY-315995, RG-13647, RWJ-38970, RWJ-46458, S-8307,S-8308, saprisartan, saralasin, Sarmesin, WK-1360, X-6803, ZD-6888,ZD-7155, ZD-8731, BIBS39, CI-996, DMP-811, DuP-532, EXP-929, L-163017,LY-301875, XH-148, XR-510, zolasartan and PD-123319.

Also included within the scope of this aspect of the invention arecombinations of the above-identified ARBs.

ARBs to be used for preparing the combination or complex in accordancewith the present invention can be purchased from commercial sources orcan be prepared according to known methods. ARBs may be used forpurposes of this invention in their free form, as well as in anysuitable salt or ester form.

Preferred salts forms include acid addition salts. The compounds havingat least one acid group (e.g., COOH or 5-tetrazolyl) can also form saltswith bases. Suitable salts with bases are, e.g., metal salts, such asalkali metal or alkaline earth metal salts, e.g., sodium, potassium,calcium or magnesium salts, or salts with ammonia or an organic amine,such as morpholine, thiomorpholine, piperidine, pyrrolidine, a mono-,di- or tri-lower alkylamine, e.g., ethyl-, tert-butyl-, diethyl-,diisopropyl-, triethyl-, tributyl- or dimethylpropylamine, or a mono-,di- or trihydroxy lower alkylamine, e.g., mono-, di- ortri-ethanolamine. Corresponding internal salts may furthermore beformed. Salts which are unsuitable for pharmaceutical uses but which canbe employed, e.g., for the isolation or purification of free compounds Ior their pharmaceutically acceptable salts, are also included. Even morepreferred salts are, e.g., selected from the mono-sodium salt inamorphous form; di-sodium salt of valsartan in amorphous or crystallineform, especially in hydrate form, thereof.

Mono-potassium salt of valsartan in amorphous form; di-potassium salt ofvalsartan in amorphous or crystalline form, especially in hydrate form,thereof.

Calcium salt of valsartan in crystalline form, especially in hydrateform, primarily the tetrahydrate thereof; magnesium salt of valsartan incrystalline form, especially in hydrate form, primarily the hexahydratethereof; calcium/magnesium mixed salt of valsartan in crystalline form,especially in hydrate form; bis-diethylammonium salt of valsartan incrystalline form, especially in hydrate form; bis-dipropylammonium saltof valsartan in crystalline form, especially in hydrate form;bis-dibutylammonium salt of valsartan in crystalline form, especially inhydrate form, primarily the hemihydrate thereof; mono-L-arginine salt ofvalsartan in amorphous form; bis-L-arginine salt of valsartan inamorphous form; mono-L-lysine salt of valsartan in amorphous form;bis-L-lysine salt of valsartan in amorphous form.

Preferably when preparing the dual-acting compound, in particular thecomplex according to the present invention, the free form of the ARB isused.

In a preferred embodiment of this invention, the angiotensin receptorblocker used in the combination or complex of the present invention isValsartan the molecular structure of which is shown below

Valsartan may be in the racemic form or as one of the two isomers shownbelow

Valsartan((S)—N-valeryl-N-{[2′-(1H-tetrazole-5-yl)-biphenyl-4-yl]-methyl}-valine)used according to the present invention can be purchased from commercialsources or can be prepared according to known methods. For example, thepreparation of valsartan is described in U.S. Pat. No. 5,399,578 and EP0 443 983, the entire disclosure of each of which is incorporated byreference herein. Valsartan may be used for purposes of this inventionin its free acid form, as well as in any suitable salt form.Additionally, esters or other derivatives of the carboxylic grouping maybe applied for the synthesis of linked pro-drugs, as well as salts andderivatives of the tetrazole grouping. Reference to ARBs includesreference to pharmaceutically acceptable salts thereof.

Preferably, the ARB is a diprotic acid. Thus, the angiotensin receptorblocker has a charge of 0, 1 or 2 depending on the pH of the solution.

In the combination of the present invention, the ARB is in the form of apharmaceutically acceptable salt selected from Na, K or NH₄, preferablyNa. This includes both the mono- and di-salt of these cations,preferably the di-salt. In particular in the case of valsartan thismeans that both the carboxylic acid moiety and the tetrazole moiety formthe salt.

In the dual-acting compound, in particular the supramolecular complex ofthe present invention, typically the free form of the ARB is employed inthe preparation and the cationic species present in the complex isintroduced by using a base, e.g. (Cat)OH.

The dual acting compound comprises a molecular moiety of a neutralendopeptidase inhibitor. This means that a molecular moiety derived froma neutral endopeptidase inhibitor is participating in the build-up ofthe dual-acting compound. The neutral endopeptidase inhibitor is part ofthe compound and connected to the ARB directly or indirectly vianon-covalent bonds. For sake of convenience, throughout the application,the term “neutral endopeptidase inhibitor” will be used when describingthis part of the compound. Neutral endopeptidase inhibitors suitable foruse in the present invention include those of formula (I)

wherein

-   -   R₂ is alkyl of 1-7 carbons, trifluoromethyl, phenyl, substituted        phenyl, —(CH₂)1 to 4-phenyl, or —(CH₂)1 to 4-substituted phenyl;    -   R₃ is hydrogen, alkyl of 1-7 carbons, phenyl, substituted        phenyl, —(CH₂)1 to 4-phenyl or —(CH₂)1 to 4-substituted phenyl;    -   R₁ is hydroxy, alkoxy of 1-7 carbons or NH₂;    -   n is an integer from 1-15;

and the term substituted phenyl refers to a substituent selected fromlower alkyl of 1-4 carbons, lower alkoxy of 1-4 carbons, lower alkylthioof 1-4 carbons, hydroxy, Cl, Br or F.

Preferred neutral endopeptidase inhibitors of formula (I) includecompounds, wherein

-   -   R₂ is benzyl;    -   R₃ is hydrogen;    -   n is an integer from 1-9; and    -   R₁ is hydroxy.

Another preferred neutral endopeptidase inhibitor is(3S,2′R)-3-{1-[2′-(ethoxycarbonyl)-4′-phenyl-butyl]-cyclopentan-1-carbonylamino}-2,3,4,5-tetrahydro-2-oxo-1H-1-benzazepine-1-aceticacid or a pharmaceutically acceptable salt thereof.

Preferred neutral endopeptidase inhibitors suitable for use in thepresent invention include, without limitation, SQ 28,603;N—[N-[1(S)-carboxyl-3-phenylpropyl]-(S)-phenylalanyl]-(S)-isoserine;N—[N—[((1 S)-carboxy-2-phenyl)ethyl]-(S)-phenylalanyl]-β-alanine;N-[2(S)-mercaptomethyl-3-(2-methylphenyl)-propionyl]methionine;(cis-4-[[[1-[2-carboxy-3-(2-methoxyethoxy)propyl]-cyclopentyl]carbonyl]amino]-cyclohexanecarboxylicacid); thiorphan; retro-thiorphan; phosphoramidon; SQ 29072;(2R,4S)-5-biphenyl-4-yl-4-(3-carboxy-propionylamino)-2-methyl-pentanoicacid ethyl ester;N-(3-carboxy-1-oxopropyl)-(4S)-p-phenylphenylmethyl)-4-amino-2R-methylbutanoicacid;(S)-cis-4-[1-[2-(5-indanyloxycarbonyl)-3-(2-methoxyethoxy)propyl]-1-cyclopentanecarboxamido]-1-cyclohexanecarboxylicacid;3-(1-[6-endo-hydroxymethylbicyclo[2,2,1]heptane-2-exo-carbamoyl]cyclopentyl)-2-(2-methoxyethyl)propanoicacid;N-(1-(3-(N-t-butoxycarbonyl-(S)-prolylamino)-2(S)-t-butoxy-carbonylpropyl)cyclopentanecarbonyl)-O-benzyl-(S)-serinemethyl ester; 4-[[2-(mercaptomethyl)-1-oxo-3-phenylpropyl]amino]benzoicacid;3-[1-(cis-4-carboxycarbonyl-cis-3-butylcyclohexyl-r-1-carbamoyl)cyclopentyl]-2S-(2-methoxyethoxymethyl)propanoicacid; N-((2S)-2-(4-biphenylmethyl)-4-carboxy-5-phenoxyvaleryl)glycine;N-(1-(N-hydroxycarbamoylmethyl)-1-cyclopentanecarbonyl)-L-phenylalanine;(S)-(2-biphenyl-4-yl)-1-(1H-tetrazol-5-yl)ethylamino) methylphosphonicacid;(S)-5-(N-(2-(phosphonomethylamino)-3-(4-biphenyl)propionyl)-2-aminoethyl)tetrazole;p-alanine;3-[1,1′-biphenyl]-4-yl-N-[diphenoxyphosphinyl)methyl]-L-alanyl;N-(2-carboxy-4-thienyl)-3-mercapto-2-benzylpropanamide;2-(2-mercaptomethyl-3-phenylpropionamido)thiazol-4-ylcarboxylic acid;(L)-(1-((2,2-dimethyl-1,3-dioxolan-4-yl)-methoxy)carbonyl)-2-phenylethyl)-L-phenylalanyl)-β-alanine;N—[N-[(L)-[1-[(2,2-dimethyl-1,3-dioxolan-4-yl)-methoxy]carbonyl]-2-phenylethyl]-L-phenylalanyl]-(R)-alanine;N—[N-[(L)-1-carboxy-2-phenylethyl]-L-phenylalanyl]-(R)-alanine;N-[2-acetylthiomethyl-3-(2-methyl-phenyl)propionyl]-methionine ethylester; N-[2-mercaptomethyl-3-(2-methylphenyl)propionyl]-methionine;N-[2(S)-mercaptomethyl-3-(2-methylphenyl)propanoyl]-(S)-isoserine;N—(S)-[3-mercapto-2-(2-methylphenyl)propionyl]-(S)-2-methoxy-(R)-alanine;N-[1-[[1(S)-benzyloxycarbonyl-3-phenylpropyl]amino]cyclopentylcarbonyl]-(S)-isoserine;N-[1-[[1(S)-carbonyl-3-phenylpropyl]amino]-cyclopentylcarbonyl]-(S)-isoserine;1,1′-[dithiobis-[2(S)-(2-methylbenzyl)-1-oxo-3,1-propanediyl]]-bis-(S)-isoserine;1,1′-[dithiobis-[2(S)-(2-methylbenzyl)-1-oxo-3,1-propanediyl]]-bis-(S)-methionine;N-(3-phenyl-2-(mercaptomethyl)-propionyl)-(S)-4-(methylmercapto)methionine;N-[2-acetylthiomethyl-3-phenyl-propionyl]-3-aminobenzoic acid;N-[2-mercaptomethyl-3-phenyl-propionyl]-3-aminobenzoic acid;N-[1-(2-carboxy-4-phenylbutyl)-cyclopentane-carbonyl]-(S)-isoserine;N-[1-(acetylthiomethyl)cyclopentane-carbonyl]-(S)-methionine ethylester;3(S)-[2-(acetylthiomethyl)-3-phenyl-propionyl]amimo-s-caprolactam;N-(2-acetylthiomethyl-3-(2-methylphenyl)propionyl)-methionine ethylester; and combinations thereof.

Neutral endopeptidase inhibitors can be purchased from commercialsources or can be prepared according to known methods, such as those setforth in any of U.S. Pat. Nos. 4,722,810, 5,223,516, 4,610,816,4,929,641, South African Patent Application 84/0670, UK 69578, U.S. Pat.No. 5,217,996, EP 00342850, GB 02218983, WO 92/14706, EP 00343911, JP06234754, EP 00361365, WO 90/09374, JP 07157459, WO 94/15908, U.S. Pat.Nos. 5,273,990, 5,294,632, 5,250,522, EP 00636621, WO 93/09101, EP00590442, WO 93/10773, U.S. Pat. No. 5,217,996, the disclosure of eachof which is incorporated by reference. Neutral endopeptidase inhibitorsmay be used for purposes of this invention in their free form, as wellas in any suitable salt form. Reference to neutral endopeptidaseinhibitors includes reference to pharmaceutically acceptable saltsthereof.

Additionally esters or other derivatives of any carboxylic grouping maybe applied for the synthesis of linked pro-drugs, as well as salts andderivatives of any other acidic grouping.

In a preferred embodiment of this invention, the NEPi is5-biphenyl-4-yl-4-(3-carboxy-priopionylamino)-2-methyl-pentanoic acidethyl ester of formula (II) or the respective hydrolysed form5-biphenyl-4-yl-4-(3-carboxy-priopionylamino)-2-methyl-pentanoic acid.

The compound of formula (II) can exist as the (2R,4S), (2R,4S), (2R,4S)or (2R,4S) isomer. Preferred is(2R,4S)-5-biphenyl-4-yl-4-(3-carboxy-propionylamino)-2-methyl-pentanoicacid ethyl ester as shown below:

The compound of formula (II) is a specific inhibitor of NEP and isdescribed in U.S. Pat. No. 5,217,996. It can be purchased fromcommercial sources or can be prepared according to known methods. Thecompound of formula (II) may be used for purposes of this invention inits free form, as well as in any suitable salt or ester form.

Preferably the NEPi is a monoprotic acid. Thus, the NEPi has a charge of0 or 1 depending on the pH of the solution.

In the combination of the present invention, the NEPi is in the form ofa pharmaceutically acceptable salt selected from Na, K or NH₄,preferably Na.

In the dual-acting compound, in particular the supramolecular complex ofthe present invention, typically the free form of the NEPi is employedin the preparation and the cationic species present in the complex isintroduced by using a base, (Cat)OH.

The dual acting compound preferably comprises non-covalent bonds betweenthe ARB and the NEPi. Alternatively or in addition, it optionallycomprises a linking moiety such as a pharmaceutically acceptable cation.

The linking moiety includes, but is not limited to, generally regardedas safe (GRAS) compounds or other pharmacologically acceptablecompounds. The linking moiety may be an ion or a neutral molecule. Inthe case wherein the linking moiety is an ion the linked pro-drug is asalt and when the linking moiety is a neutral molecule the linkedpro-drug is a co-crystal. Without being bound by any particular theory,the acidic portion of the ARB and NEPi donate a proton to the basiclinking moiety such that all three components then become united to formone molecule. When the linked pro-drug is ingested by the subjectintended to be treated the more acidic nature of the ingestionenvironment causes the linked pro-drug to separate into individualcomponents concomitant with ingestion and absorption and therefore beconverted into active agents to provide their beneficial biologicalaction to treat the intended diseases.

In the case of a linked pro-drug salt or the dual-acting compound, thelinking moiety or the cation, respectively, is preferably a positivelycharged mono-, di- or tri-valent cation, an organic base or an aminoacid. Preferred cations (Cat) both for the linked pro-drug in generaland the dual-acting compound, in particular the complex are basiccations, even more preferably metallic cations. Preferred metalliccations include, but are not limited to Na, K, Ca, Mg, Zn, Fe or NH₄.Amine bases and salt forming agents may also be employed, such asbenzathine, hydrabamine, ethylenediamine, n-n-dibenzyl-ethylenediamine,L-arginine, choline hydroxide, N-methyl-glucamine, (Meglumine),L-Lysine, dimethylaminoethanol (Deanol), t-butylamine, diethylamine,2-(diethylamino)-ethanol, 4-(2-hydroxyethyl)-morpholine, Thromethanine(TRIS), 4-acetamidophenol, 2-amino-2-methyl-1,3-propanediol,2-amino-2-methyl-propanol, benzylamine, cyclohexylamine, diethanolamine,ethanolamine, imidazole, piperazine and triethanolamine.

Most preferably, the cation is Na, K or NH₄, such as Na. In oneembodiment Ca is preferred.

In the case of a linked pro-drug co-crystal, the linking moiety is mayalso be a neutral molecule which provides hydrogen-bondingfunctionality.

In one embodiment, the linked pro-drugs of this invention arerepresented as set forth below, wherein scheme (1) and (2) represent asalt and scheme (3) represents a co-crystal:

NEPi·Xa·ARB  scheme (1)

NEPi·XaYb·ARB  scheme (2)

NEPi·Zc·ARB  scheme (3),

wherein

-   -   X is Ca, Mg, Zn or Fe;    -   Y is Na, K or NH₄;    -   Z is a neutral molecule; and    -   a, b and c reflect the stoichiometry of the linked pro-drug,        preferably, a, b and c are a valence of 1⁺, 2⁺ or 3⁺.

For the linked pro-drugs of schemes (1) and (2), above, preferably theNEPi is a monoprotic acid and ARB is a diprotic acid. The angiotensinreceptor blocker has a charge of 0, 1 or 2 and the NEPi has a charge of0 or 1 depending on the pH of the solution, while the overall moleculewill be neutral. Ratios of ARB to NEPi will be 1:1, 1:2, 1:3, 3:1, 2:1,1:1, preferably 1:1, 1:2 or 1:3, most preferably 1:1.

Multi-component salts, particularly with zinc and calcium have beenreported in the literature, e.g., Chem Pharm Bull, Vol. 53, p. 654(2005). These ions require a coordination geometry that facilitates thecrystallization of multi-component systems. The metal ions havecoordinating geometries governed by the atomic orbitals for each species

Valsartan comprises two acidic groupings: the carboxylic acid and thetetrazole. In one embodiment of this aspect of the present invention,the molecular structure of linked pro-drugs of valsartan and a NEPicomprise a linkage between the carboxylic acid and the linking moiety ora linkage between the tetrazole grouping and the linking moiety. In yetanother embodiment, the linked pro-drug comprises a trivalent linkingmoiety linked to the valsartan carboxylic acid grouping, the tetrazolegrouping and the NEPi grouping.

In an embodiment of this aspect of the invention, valsartan is linked to(2R,4S)-5-biphenyl-4-yl-4-(3-carboxy-propionylamino)-2-methyl-pentanoicacid ethyl ester by a calcium salt ion.

In a preferred embodiment of the present application, the angiotensinreceptor antagonist and the neutral endopeptidase inhibitor are presentin a molar ratio of 1:1, 1:2, 1:3, 3:1, 2:1, more preferably 1:1 in thecombination as well as in the supramolecular complex. This is also truefor the linked pro-drug. Moreover, in the complex, angiotensin receptorantagonist, the neutral endopeptidase inhibitor and the cation arepresent in a molar ratio of 1:1:1, 1:1:2, 1:1:3, more preferably 1:1:3.This applies equally to the linked pro-drug.

The combination or the dual-acting compound, in particular the complexof the present invention may contain a solvent. This is particularlypreferred in the case of the dual-acting compound, in particular thecomplex, where the solvent may contribute to the intermolecularstructure, e.g. the supramolecular interactions. Preferred solventsinclude water, methanol, ethanol, 2-propanol, acetone, ethyl acetate,methyl-t-butylether, acetonitrile, toluene, and methylene chloride,preferably water. If a solvent is present, one or more molecules permolecule of the active agent can be present. In this case, namely if astoichiometric amount of the solvent is present, preferably 1, 2, 3, 4or 5, more preferably 3, molecules of solvent, such as water, can bepresent per molecule of active agent. Alternatively, the solvent may bepresent in non-stoichiometric amounts. This means preferably anystoichiometric fraction of the solvent, such as 0.25, 0.5, 0.75, 1.25,1.5, 1.75, 2.25, 2.5, 2.75, 3.25, 3.5, 3.75, 4.25, 4.5 and 4.75,preferably 2.5, molecules of solvent, such as water, can be present permolecule of active agent. If the dual-acting compound, in particular thecomplex is in the crystalline form, the solvent may be part of themolecular packing and be trapped in the crystal lattice.

Thus in a preferred embodiment of the present invention, the dual-actingcompound, in particular the supramolecular complex is described by thesum formula:

[ARB(NEPi)]Na₃.xH₂O, wherein x is 0, 1, 2 or 3, such as 3, preferably

[ARB(NEPi)]Na₃.xH₂O, wherein x is 0, 1, 2 or 3, such as 3, morepreferably

[valsartan((2R,4S)-5-biphenyl-4-yl-4-(3-carboxy-propionylamino)-2-methyl-pentanoicacid ethyl ester)]Na₃.x H₂O, wherein x is 0, 1, 2 or 3, such as 3.

Thus in a preferred embodiment of the present invention, the dual-actingcompound, in particular the supramolecular complex is described by thesum formula:

[ARB(NEPi)]Na₁₃.xH₂O, wherein x is 0 to 3, such as 2.5, preferably

[ARB(NEPi)]Na₃.xH₂O, wherein x is 0 to 3, such as 2.5, more preferably

[(N-valeryl-N-{[2′-(1H-tetrazole-5-yl)-biphenyl-4-yl]-methyl}-valine)(5-biphenyl-4-yl-4-(3-carboxy-priopionylamino)-2-methyl-pentanoic acidethyl ester]Na₃.xH₂O, in particular[((S)—N-valeryl-N-{[2′-(1H-tetrazole-5-yl)-biphenyl-4-yl]-methyl}-valine)((2R,4S)-5-biphenyl-4-yl-4-(3-carboxy-propionylamino)-2-methyl-pentanoicacid ethyl ester]Na₃.xH₂O, wherein x is 0 to 3, such as 2.5. In thismost preferred example, the complex is termed trisodium[3-((1S,3R)-1-biphenyl-4-ylmethyl-3-ethoxycarbonyl-1-butylcarbamoyl)propionate-(S)-3′-methyl-2′-(pentanoyl{2″-(tetrazol-5-ylate)biphenyl-4′-ylmethyl}amino)butyrate]hemipentahydrate.

A simplified structure of trisodium [3-((1S,3R)-1-biphenyl-4-ylmethyl-3-ethoxycarbonyl-1-butylcarbamoyl)propionate-(S)-3′-methyl-2′-(pentanoyl{2″-(tetrazol-5-ylate)biphenyl-4′-ylmethyl}amino)butyrate]hemipentahydrate used to formally calculate the relative molecular mass,is shown below.

Valsartan comprises two acidic groupings: the carboxylic acid and thetetrazole. In one embodiment of this aspect of the present invention,the molecular structure of the dual-acting compound, in particular, thecomplex, of valsartan and a NEPi comprises an interaction between thecarboxylic acid and the cation, such as Na, or the solvent, such aswater, or a linkage between the tetrazole grouping and the cation, suchas Na, or the solvent, such as water. In yet another embodiment, thedual-acting compound, in particular, the complex, comprises aninteraction between the valsartan carboxylic acid grouping, thetetrazole grouping or the NEPi grouping and the cation, such as Na, orthe solvent, such as water.

The combination or dual-acting compound, in particular, the complex, ofthe present invention is preferably in the solid form. In the solidstate it can be in the crystalline, partially crystalline, amorphous, orpolymorphous form, preferably in the crystalline form.

The dual-acting compound, in particular, the complex, of the presentinvention is distinct from a combination of an ARB and a NEPi obtainedby simply physically mixing the two active agents. Thus, it can havedifferent properties that make it particularly useful for manufacturingand therapeutic applications. The difference of the dual-actingcompound, in particular, the complex, and the combination can beexemplified by the dual-acting compound of(S)—N-valeryl-N-{[2′-(1H-tetrazole-5-yl)-biphenyl-4-yl]-methyl}-valineand(2R,4S)-5-biphenyl-4-yl-4-(3-carboxy-propionylamino)-2-methyl-pentanoicacid ethyl ester which is characterized by very distinct spectral peaksand shifts that are not observed in the physical mixture.

Specifically, such a dual-acting compound is preferably characterized byan X-ray powder diffraction pattern taken with a Scintag XDS2000 powderdiffractometer using Cu-Ka radiation (lamda=1.54056 A) with aPeltier-cooled Silicon detector at room temperature (25 degree C.). Scanrange was from 1.5 degree to 40 degree in 2 theta with a scan rate of 3degree/minute. The most important reflections in the X-ray diffractiondiagram comprise the following interlattice plane intervals:

The preferred characterization of trisodium[3-((1S,3R)-1-biphenyl-4-ylmethyl-3-ethoxycarbonyl-1-butylcarbamoyl)propionate-(S)-3′-methyl-2′-(pentanoyl{2″-(tetrazol-5-ylate)biphenyl-4′-ylmethyl}amino)butyrate]hemipentahydrate is obtained from the interlattice plane intervals d ofthe ascertained X-ray diffraction diagrams, whereby, in the following,average values 2Θ in [°] are indicated (error limit of ±0.2)

4.5, 5.5, 5.6, 9.9, 12.8, 15.7, 17.0, 17.1, 17.2, 18.3, 18.5, 19.8,21.5, 21.7, 23.2, 23.3, 24.9, 25.3, 27.4, 27.9, 28.0, 30.2.

or with an error limit of 0.1:

4.45, 5.52, 5.57, 9.94, 12.82, 15.66, 17.01, 17.12, 17.2, 18.32, 18.46,19.76, 21.53, 21.72, 23.17, 23.27, 24.88, 25.3, 27.4, 27.88, 28.04,30.2.

The most intensive reflections in the X-ray diffraction pattern show thefollowing interlattice plane intervals:

2Θ in [°]: 4.5, 5.6, 12.8, 17.0, 17.2, 19.8, 21.5, 27.4, in particular4.45, 5.57, 17.01, 17.2, 19.76, 21, 27.4.

A preferred method of checking the above-indicated average values of theinterlattice plane intervals and intensities measured by experimentationfrom X-ray diffraction, for a given substance, consists in calculatingthese intervals and their intensities from the comprehensive singlecrystal structure determination. This structure determination yieldscell constants and atom positions, which enable the X-ray diffractiondiagram corresponding to the solid to be calculated by means ofcomputer-aided calculation methods. The program used is Powder Patternwithin the application software Materials Studio (Accelrys). Acomparison of these data, namely the interlattice plane intervals andintensities of the most important lines of trisodium [3-((1S,3R)-1-biphenyl-4-ylmethyl-3-ethoxycarbonyl-1-butylcarbamoyl)propionate-(S)-3′-methyl-2′-(pentanoyl{2″-(tetrazol-5-ylate)biphenyl-4′-ylmethyl}amino)butyrate]hemipentahydrate, obtained from measurements and from calculating thesingle crystal data, is illustrated in the table below.

TABLE measured calculated measured calculated 2θ in Inten- 2θ in Inten-2θ in Inten- 2θ in Inten- [°] sity [°] sity [°] sity [°] sity 4.45 very4.15 very 19.76 strong 19.6 very strong strong weak 5.52 Strong 5 strong21.53 weak 19.8 very weak 5.57 strong 6.5 strong 21.72 very 21.4 veryweak weak 9.94 very 9.75 weak 23.17 weak 23.1 very weak weak 12.82 very12.6 weak 23.27 weak 23.15 very strong weak 15.66 very 15.05 strong24.88 very very weak weak weak 17.01 weak 16.9 very 25.3 weak 25.3 verystrong weak 17.12 strong 17.1 strong 27.4 weak 27.3 very weak 17.2 weak17.15 weak 27.88 very 27.9 very weak weak 18.32 weak 18.25 very 28.04weak weak 18.46 weak 18.3 weak 30.2 weak Relative intensity between 100%to 50% is referred to as very strong, 50% to 10% as strong, 10% to 5% asweak, and below 5% as very weak.

The invention relates to trisodium [3-((1S,3R)-1-biphenyl-4-ylmethyl-3-ethoxycarbonyl-1-butylcarbamoyl)propionate-(S)-3′-methyl-2′-(pentanoyl{2″-(tetrazol-5-ylate)biphenyl-4′-ylmethyl}amino)butyrate] hemipentahydrate, a crystallinesolid which is characterized by the data and parameters obtained fromsingle crystal X-ray analysis and X-ray powder patterns. An in-depthdiscussion of the theory of the methods of single crystal X-raydiffraction and the definition of the evaluated crystal data and theparameters may be found in Stout & Jensen, X-Ray StructureDetermination; A Practical Guide, Mac Millian Co., New York, N.Y. (1968)chapter 3.

Crystal data sum formula C₄₈H₅₅N₆O₈Na₃•2.5H₂O molecular mass 957.99crystal colour colourless crystal shape tabular: hexagonal crystalsystem monoclinic space group P2₁ Cell parameters a = 20.344 Å b =42.018 Å c = 20.374 Å α = 90° β = 119.29° γ = 90° volume of unit cell15190.03 Å³ Z (the number of asymmetric units 2 in the unit cell)calculated density 1.26845 g/cm3 Single crystal X-ray measurement datadiffractometer Nonius KappaCCD X-ray generator Nonius FR571 X-raygenerator with a copper rotating anode temperature 270 K and 150 KNotes: Two data sets on two suitable single crystals were collected attwo different temperatures to assure no phase change during cooling.None of the hydrogen atoms on the water or amine nitrogen atoms wereobserved in the Fourier maps so they were not included in therefinement.

Computer Program Used to Solve the Structure

SHELXD (Sheldrick, Göttingen)

In three dimensions, the unit cell is defined by three edge lengths a,b, and c, and three interaxial angles α, β, und γ. In this way, thevolume of the unit cell V_(c) is determined. A differentiateddescription of these crystal parameters is illustrated in chapter 3 ofStout & Jensen (see above). The details for trisodium [3-((1S,3R)-1-biphenyl-4-ylmethyl-3-ethoxycarbonyl-1-butylcarbamoyl)propionate-(S)-3′-methyl-2′-(pentanoyl{2″-(tetrazol-5-ylate)biphenyl-4′-ylmethyl}amino)butyrate]hemipentahydrate from the single crystal measurements, especially theatom coordinates, the isotropic thermal parameters, the coordinates ofthe hydrogen atoms as well as the corresponding isotropic thermalparameters, show that a monoclinic unit cell exists, its cell content oftwelve formula units of C₄₈H₅₅N₆O₈Na₃.2.5 H₂O occurring as a result oftwo asymmetric units on two-fold positions.

The acentric space group P2₁ determined from the single crystal X-raystructure is a common space group for enantiomorphically pure molecules.In this space group there are two general positions which means that fortwelve formula units in the unit cell there must be 18 sodium ions and15 waters in the asymmetric unit.

A pictorial representation of the unit cell of the supramolecularcomplex of trisodium[3-((1S,3R)-1-biphenyl-4-ylmethyl-3-ethoxycarbonyl-1-butylcarbamoyl)propionate-(S)-3′-methyl-2′-(pentanoyl{2″-(tetrazol-5-ylate)biphenyl-4′-ylmethyl}amino)butyrate]hemipentahydrate comprising two asymmetric units is shown in FIG. 1.

Based on the single crystal structure solution, the asymmetric unit ofthe trisodium[3-((1S,3R)-1-biphenyl-4-ylmethyl-3-ethoxycarbonyl-1-butylcarbamoyl)propionate-(S)-3′-methyl-2′-(pentanoyl{2″-(tetrazol-5-ylate)biphenyl-4′-ylmethyl}amino)butyrate]hemipentahydrate supramolecule comprises six each of ARB and NEPimoieties, 18 sodium atoms, and 15 water molecules. Trisodium [3-((1S,3R)-1-biphenyl-4-ylmethyl-3-ethoxycarbonyl-1-utylcarbamoyl)propionate-(S)-3′-methyl-2′-(pentanoyl{2″-(tetrazol-5-ylate)biphenyl-4′-ylmethyl}amino)butyrate]hemipentahydrate may be considered a sodium supramolecular complex,coordinated by oxygen ligands. These oxygens come from twelvecarboxylate groups and eighteen carbonyl groups of the above moieties,and from 13 of the 15 water molecules. The crystal is an infinite3-dimensional network of these sodium complexes.

Such a compound may also be characterized by an infrared absorptionspectrum obtained using Attenuated Total Reflection Fourier TransformInfrared (ATR-FTIR) spectrometer (Nicolet Magna-IR 560) showing thefollowing significant bands, expressed in reciprocal wave numbers(cm⁻¹):

2956 (w), 1711 (st), 1637 (st), 1597 (st), 1488 (w), 1459 (m), 1401(st), 1357 (w), 1295 (m), 1266 (m), 1176 (w), 1085 (m), 1010 (w),942(w), 907 (w), 862 (w), 763 (st), 742 (m), 698 (m), 533 (st).Characteristic to the complex are in particular the following peaks1711(st), 1637(st), 1597(st) and 1401(st). The error margin for allabsorption bands of ATR-IR is ±2 cm⁻¹. The intensities of the absorptionbands are indicated as follows: (w)=weak; (m)=medium; and (st)=strongintensity.

Such a compound may also be characterized by a Raman spectrum measuredby dispersive Raman spectrometer with 785 nm laser excitation source(Kaiser Optical Systems, Inc.) showing the following significant bandsexpressed in reciprocal wave numbers (cm⁻¹):

3061 (m), 2930 (m, broad), 1612 (st), 1523 (m), 1461 (w), 1427 (w), 1287(st), 1195 (w), 1108 (w), 11053 (w), 1041 (w), 1011 (w), 997 (m), 866(w), 850 (w), 822 (w), 808 (w), 735 (w), 715 (w), 669 (w), 643 (w), 631(w), 618 (w), 602 (w), 557 (w), 522 (w), 453 (w), 410 (w), 328 (w).

The error margin for all Raman bands is ±2 cm⁻¹. The intensities of theabsorption bands are indicated as follows: (w)=weak; (m)=medium; and(st)=strong intensity.

Such a compound may also be characterized by distinct melting propertiesmeasured by differential scanning calorimetry (DSC). Using Q1000 (TAInstruments) instrument, the melting onset temperature and the peakmaximum temperature for such a complex are observed at 139° C. and 145°C., respectively. The heating rate is 10 K/min.

The second embodiment of the present invention is directed topharmaceutical compositions comprising a combination, a linked pro-drugor a dual-acting compound, in particular the complex as described hereinand at least one pharmaceutically acceptable additive. The detailsregarding the combination and the complex, including the ARB and theNEPi, are as described above with regard to the first embodiment of theinvention.

The pharmaceutical compositions according to the invention can beprepared in a manner known per se and are those suitable for enteral,such as oral or rectal, and parenteral administration to mammals(warm-blooded animals), including man, comprising a therapeuticallyeffective amount of the combination or dual-acting compound, inparticular the complex, alone or in combination with at least onepharmaceutically acceptable carrier, especially suitable for enteral orparenteral application. Typical oral formulations include tablets,capsules, syrups, elixirs and suspensions. Typical injectableformulations include solutions and suspensions.

Pharmaceutically acceptable additives suitable for use in the presentinvention include, without limitation and provided they are chemicallyinert so that they do not adversely affect the combination or thedual-acting compound, in particular the complex of the presentinvention, diluents or fillers, disintegrants, glidants, lubricants,binders, colorants and combinations thereof. The amount of each additivein a solid dosage formulation may vary within ranges conventional in theart. Typical pharmaceutically acceptable carriers for use in theformulations described above are exemplified by: sugars, such aslactose, sucrose, mannitol and sorbitol; starches, such as cornstarch,tapioca starch and potato starch; cellulose and derivatives, such assodium carboxymethyl cellulose, ethyl cellulose and methyl cellulose;calcium phosphates, such as dicalcium phosphate and tricalciumphosphate; sodium sulfate; calcium sulfate; polyvinylpyrrolidone;polyvinyl alcohol; stearic acid; alkaline earth metal stearates, such asmagnesium stearate and calcium stearate; stearic acid; vegetable oils,such as peanut oil, cottonseed oil, sesame oil, olive oil and corn oil;non-ionic, cationic and anionic surfactants; ethylene glycol polymers;p-cyclodextrin; fatty alcohols; and hydrolyzed cereal solids, as well asother non-toxic compatible fillers, binders, disintegrants, buffers,preservatives, antioxidants, lubricants, flavoring agents and the likecommonly used in pharmaceutical formulations.

Pharmaceutical preparations for enteral or parenteral administrationare, e.g., in unit dose forms, such as coated tablets, tablets, capsulesor suppositories and also ampoules. These are prepared in a manner whichis known per se, e.g., using conventional mixing, granulation, coating,solubilizing or lyophilizing processes. Thus, pharmaceuticalcompositions for oral use can be obtained by combining the linkedpro-drug, combination or dual-acting compound, in particular the complexwith solid excipients, if desired, granulating a mixture which has beenobtained, and, if required or necessary, processing the mixture orgranulate into tablets or coated tablet cores after having addedsuitable auxiliary substances.

The dosage of the active compounds in the combination or dual-actingcompound, in particular the complex can depend on a variety of factors,such as mode of administration, homeothermic species, age and/orindividual condition. The projected efficacy in animal disease modelsranges from about 0.1 mg/kg/day to about 1000 mg/kg/day given orally,and the projected dose for human treatment ranges from about 0.1 mg/dayto about 2000 mg/day. Preferred ranges are from about 40 mg/day to about960 mg/day of the linked pro-drug, preferably about 80 mg/day to about640 mg/day. The ARB component is administered in a dosage of from about40 mg/day to about 320 mg/day and the NEPi component is administered ina dosage of from about 40 mg/day to about 320 mg/day. More specifically,the dosages of ARB/NEPi, respectively, include 40 mg/40 mg, 80 mg/80 mg,160 mg/160 mg, 320 mg/320 mg, 40 mg/80 mg, 80 mg/160 mg, 160 mg/320 mg,320 mg/640 mg, 80 mg/40 mg, 160 mg/80 mg and 320 mg/160 mg,respectively. These dosages are “therapeutically effective amounts”.Preferred dosages for the linked pro-drug, combination or dual-actingcompound, in particular the complex of the pharmaceutical compositionaccording to the present invention are therapeutically effectivedosages.

The pharmaceutical compositions may contain in addition anothertherapeutic agent, e.g., each at an effective therapeutic dose asreported in the art. Such therapeutic agents include:

a) antidiabetic agents such as insulin, insulin derivatives andmimetics; insulin secretagogues such as the sulfonylureas, e.g.,Glipizide, glyburide and Amaryl; insulinotropic sulfonylurea receptorligands such as meglitinides, e.g., nateglinide and repaglinide;peroxisome proliferator-activated receptor (PPAR) ligands; proteintyrosine phosphatase-1B (PTP-1B) inhibitors such as PTP-112; GSK3(glycogen synthase kinase-3) inhibitors such as SB-517955, SB-4195052,SB-216763, NN-57-05441 and NN-57-05445; RXR ligands such as GW-0791 andAGN-194204; sodium-dependent glucose cotransporter inhibitors such asT-1095; glycogen phosphorylase A inhibitors such as BAY R3401;biguanides such as met-formin; alpha-glucosidase inhibitors such asacarbose; GLP-1 (glucagon like peptide-1), GLP-1 analogs such asExendin-4 and GLP-1 mimetics; and DPPIV (dipeptidyl peptidase IV)inhibitors such as LAF237;

b) hypolipidemic agents such as 3-hydroxy-3-methyl-glutaryl coenzyme A(HMG-CoA) reductase inhibitors, e.g., lovastatin, pitavastatin,simvastatin, pravastatin, cerivastatin, mevastatin, velostatin,fluvastatin, dalvastatin, atorvastatin, rosuvastatin and rivastatin;squalene synthase inhibitors; FXR (farnesoid X receptor) and LXR (liverX receptor) ligands; cholestyramine; fibrates; nicotinic acid andaspirin;

c) anti-obesity agents such as orlistat; and

d) anti-hypertensive agents, e.g., loop diuretics such as ethacrynicacid, furosemide and torsemide; angiotensin converting enzyme (ACE)inhibitors such as benazepril, captopril, enalapril, fosinopril,lisinopril, moexipril, perinodopril, quinapril, ramipril andtrandolapril; inhibitors of the Na-K-ATPase membrane pump such asdigoxin; ACE/NEP inhibitors such as omapatrilat, sampatrilat andfasidotril; β-adrenergic receptor blockers such as acebutolol, atenolol,betaxolol, bisoprolol, metoprolol, nadolol, propranolol, sotalol andtimolol; inotropic agents such as digoxin, dobutamine and milrinone;calcium channel blockers such as amlodipine, bepridil, diltiazem,felodipine, nicardipine, nimodipine, nifedipine, nisoldipine andverapamil; aldosterone receptor antagonists; and aldosterone synthaseinhibitors. Most preferred combination partners are diuretics, such ashydrochlorothiazide, and/or calcium channel blockers, such as amlodipineor a salt thereof.

Other specific anti-diabetic compounds are described by Patel Mona inExpert Opin Investig Drugs, 2003, 12(4), 623-633, in the FIGS. 1 to 7,which are herein incorporated by reference. A compound of the presentinvention may be administered either simultaneously, before or after theother active ingredient, either separately by the same or differentroute of administration or together in the same pharmaceuticalformulation.

The structure of the therapeutic agents identified by code numbers,generic or trade names may be taken from the actual edition of thestandard compendium “The Merck Index” or from databases, e.g., PatentsInternational (e.g. IMS World Publications). The corresponding contentthereof is hereby incorporated by reference.

Accordingly, the present invention provides pharmaceutical compositionsin addition a therapeutically effective amount of another therapeuticagent, preferably selected from antidiabetics, hypolipidemic agents,anti-obesity agents or anti-hypertensive agents, most preferably fromantidiabetics, anti-hypertensive agents or hypolipidemic agents asdescribed above.

The person skilled in the pertinent art is fully enabled to select arelevant test model to prove the efficacy of a combination of thepresent invention in the hereinbefore and hereinafter indicatedtherapeutic indications.

Representative studies are carried out with trisodium [3-((1S,3R)-1-biphenyl-4-ylmethyl-3-ethoxycarbonyl-1-butylcarbamoyl)propionate-(S)-3′-methyl-2′-(pentanoyl{2″-(tetrazol-5-ylate)biphenyl-4′-ylmethyl}amino)butyrate]hemipentahydrate, e.g. applying the following methodology:

The antihypertensive and neutral endopeptidase 24.11 (NEP)-inhibitoryactivities of trisodium [3-((1S,3R)-1-biphenyl-4-ylmethyl-3-ethoxycarbonyl-1-butylcarbamoyl)propionate-(S)-3′-methyl-2′-(pentanoyl{2″-(tetrazol-5-ylate)biphenyl-4′-ylmethyl}amino)butyrate]hemipentahydrateis assessed in conscious rats. The blood pressure-lowering effect isevaluated in double-transgenic rats (dTGRs) that overexpress both humanrenin and its substrate, human angiotensinogen (Bohlender, et al, Highhuman renin hypertension in transgenic rats. Hypertension; 29(1 Pt2):428-34, 1997). Consequently, these animals exhibit an angiotensinII-dependent hypertension. The NEP-inhibitory effect of trisodium[3-((1S,3R)-1-biphenyl-4-ylmethyl-3-ethoxycarbonyl-1-butylcarbamoyl)propionate-(S)-3′-methyl-2′-(pentanoyl{2″-(tetrazol-5-ylate)biphenyl-4′-ylmethyl}amino)butyrate]hemipentahydrate is determined in conscious Sprague-Dawley rats infusedwith exogenous atrial natriuretic peptide (ANP). Potentiation of plasmaANP levels is used as an index of NEP inhibition in vivo. In bothmodels, trisodium[3-((1S,3R)-1-biphenyl-4-ylmethyl-3-ethoxycarbonyl-1-butylcarbamoyl)propionate-(S)-3′-methyl-2′-(pentanoyl{2″-(tetrazol-5-ylate)biphenyl-4′-ylmethyl}amino)butyrate]hemipentahydrate is administered orally as a powder in gelatin minicapsules. The results are summarized below.

-   -   Trisodium        [3-((1S,3R)-1-biphenyl-4-ylmethyl-3-ethoxycarbonyl-1-butylcarbamoyl)propionate-(S)-3′-methyl-2′-(pentanoyl{2″-(tetrazol-5-ylate)biphenyl-4′-ylmethyl}amino)butyrate]        hemipentahydrate exhibits a dose-dependent and long-lasting        antihypertensive effect after oral administration in conscious        dTGRs, a rat model of fulminant hypertension.    -   Oral administration of trisodium        [3-((1S,3R)-1-biphenyl-4-ylmethyl-3-ethoxycarbonyl-1-butylcarbamoyl)propionate-(S)-3′-methyl-2′-(pentanoyl{2″-(tetrazol-5-ylate)biphenyl-4′-ylmethyl}amino)butyrate]        hemipentahydrate rapidly and dose-dependently inhibits NEP with        a long duration of action, as reflected by its potentiation of        plasma ANP immunoreactivity (ANPir) in conscious Sprague-Dawley        rats infused with exogenous ANP.

Antihypertensive Effect In Vivo

The dTGRs are instrumented with radiotelemetry transmitters forcontinuous measurement of arterial blood pressure and heart rate.Animals are randomly assigned to vehicle (empty capsule) or treatment(at 2, 6, 20 or 60 mg/kg, p.o.) groups. Baseline 24-hr mean arterialpressure (MAP) is approximately 170-180 mmHg in all groups. Trisodium[3-((1S,3R)-1-biphenyl-4-ylmethyl-3-ethoxycarbonyl-1-butylcarbamoyl)propionate-(S)-3′-methyl-2′-(pentanoyl{2″-(tetrazol-5-ylate)biphenyl-4′-ylmethyl}amino)butyrate]hemipentahydrate dose-dependently reduces MAP. The values obtained fromthe treatment groups are dose-dependent, and the results from the threehighest doses are significantly different from the vehicle controls

Inhibition of NEP In Vivo

The extent and duration of trisodium[3-((1S,3R)-1-biphenyl-4-ylmethyl-3-ethoxycarbonyl-1-butylcarbamoyl)propionate-(S)-3′-methyl-2′-(pentanoyl{2″-(tetrazol-5-ylate)biphenyl-4′-ylmethyl}amino)butyrate]hemipentahydrate for NEP inhibition in vivo is assessed withmethodologies as described previously (Trapani, et al, CGS 35601 and itsorally active prodrug CGS 37808 as triple inhibitors ofendothelin-converting enzyme-1, neutral endopeptidase 24.11, andangiotensin-converting enzyme. J Cardiovasc Pharmacol; 44(Suppl1):S211-5, 2004). Rat ANP(1-28) is infused intravenously at a rate of450 ng/kg/min in conscious, chronically cannulated, male Sprague-Dawleyrats. After one hour of infusion, rats are randomly assigned to one ofsix groups: untreated control, vehicle (empty capsule) control, or oneof four doses of drug (2, 6, 20, or 60 mg/kg, p.o.). ANP infusion iscontinued for an additional eight hours. Blood samples are collected formeasuring plasma ANPir by a commercial enzyme immunoassay kit at −60 min(i.e., before initiating ANP infusion), −30 min (after 30 min of ANPinfusion), 0 min (“baseline”; after 60 min of ANP infusion but beforedosing with drug or its vehicle), and at 0.25, 0.5, 1, 2, 3, 4, 5, 6, 7,and 8 hr post-dosing.

Before ANP infusion, ANPir is low (0.9-1.4 ng/ml) and similar in all sixgroups. ANP infusion rapidly (by 30 min) elevates ANPir to ˜10 ng/ml.This ANPir level is sustained for the duration of the experiment in theuntreated and vehicle control groups. In contrast, trisodium[3-((1S,3R)-1-biphenyl-4-ylmethyl-3-ethoxycarbonyl-1-butylcarbamoyl)propionate-(S)-3′-methyl-2′-(pentanoyl{2″-(tetrazol-5-ylate)biphenyl-4′-ylmethyl}amino)butyrate]hemipentahydraterapidly (within 15 min) and dose-dependently augments ANPir. In summary,orally administered LCZ696 rapidly and dose-dependently inhibited NEPwith a long duration of action as reflected by the potentiation ofplasma ANPir.

The available results indicate an unexpected therapeutic effect of acompound according to the invention.

In a third aspect, the present invention is directed to a method ofmaking a linked pro-drug of an ARB or a pharmaceutically acceptable saltthereof and a NEPi or a pharmaceutically acceptable salt thereofcomprising the steps of:

-   -   (a) adding an inorganic salt forming agent to a solvent to form        a linked pro-drug salt forming solution;    -   (b) adding the salt forming solution to a mixture of an ARB and        a NEPi such that the ARB and NEPi form a linked pro-drug; and    -   (c) isolating the linked pro-drug.

Preferably, the components are added in an equivalent amount.

The inorganic salt forming agent includes, but is not limited to,calcium hydroxide, zinc hydroxide, calcium methoxide, calcium acetate,calcium hydrogen carbonate, calcium formate, magnesium hydroxide,magnesium acetate, magnesium formate and magnesium hydrogen carbonate,sodium hydroxide, sodium methoxide, sodium acetate, sodium formate. Theinorganic salt forming agent releases the linking moiety into thesolvent such that when an ARB and a NEPi are present a linked pro-drugis formed.

Solvents included in the scope of the present invention include, but arenot limited to, solvents in which the ARB, NEPi and inorganic saltforming agent preferably exhibit a lower solubility that allows thelinked pro-drug to crystallize. Such solvents may comprise, but are notlimited to, water, methanol, ethanol, 2-propanol, ethyl acetate,methyl-t-butylether, acetonitrile, toluene, and methylene chloride andmixtures of such solvents.

The inorganic salt forming agent and the solvent when combined shouldhave a pH which promotes linked pro-drug formation. The pH may bebetween about 2 and about 6, preferably between about 3 and about 5,most preferably between 3.9 and 4.7.

The linked pro-drug is isolated by crystallization and chromatography.Specific types of chromatography include, e.g., ligand specific resinchromatography, reverse phase resin chromatography and ion-exchangeresin chromatography.

A specific example comprises contacting a divalent salt of one componentwith a monovalent salt of the other component of the linked pro-drug.Specifically the mixed salt of valsartan and a mono-basic NEPi aresynthesized by contacting the calcium salt of valsartan with the sodiumsalt of the NEPi component. Isolation of the desired mixed salt iscarried out by selective crystallization or chromatography using ligandspecific resins, reverse phase resins or ion-exchange resins. Similarlythis process can be conducted with a monovalent salt of both components,such as the sodium salt of both components.

In another embodiment of this aspect of the invention, a co-crystal ofthe linked pro-drug is obtained. In a method of making a linked pro-drugco-crystal the inorganic salt forming agent is replaced with a neutralmolecule which provides hydrogen binding properties. The solvent may bepart of the molecular packing and be trapped in the crystal lattice.

In a preferred embodiment of the third aspect, the present invention isdirected to a method of preparing a dual-acting compound comprising

-   -   (a) an angiotensin receptor antagonist;    -   (b) a neutral endopeptidase inhibitor (NEPi); and optionally    -   (c) a pharmaceutically acceptable cation;    -   said method comprising the steps of:    -   (i) dissolving an angiotensin receptor antagonist and a neutral        endopeptidase inhibitor (NEPi) in a suitable solvent;    -   (ii) dissolving a basic compound of Cat in a suitable solvent,        wherein Cat is a cation;    -   (iii) combining the solutions obtained in steps (i) and (ii);    -   (iv) precipitation of the solid, and drying same to obtain the        dual-acting compound; or alternatively    -   obtaining the dual-acting compound by exchanging the solvent(s)        employed in steps (i) and (ii) by    -   (iva) evaporating the resulting solution to dryness;    -   (va) re-dissolving the solid in a suitable solvent;    -   (via) precipitation of the solid and drying same to obtain the        dual-acting compound.

The details regarding the complex, including the ARB, the NEPi and thecation, are as described above with regard to the first embodiment ofthe invention.

Preferably, in step (i) the ARB and the NEPi are added in an equivalentmolar amount. Both the ARB and the NEPi are preferably used in the freeform. The solvent used in step (i) may be any solvent that allowsdissolution of both the ARB and the NEPi. Preferred solvents includethose mentioned above, namely water, methanol, ethanol, 2-propanol,acetone, ethyl acetate, isopropyl acetate, methyl-t-butylether,acetonitrile, toluene, DMF, NMF and methylene chloride and mixtures ofsuch solvents, such as ethanol-water, methanol-water, 2-propanol-water,acetonitrile-water, acetone-water, 2-propanol-toluene, ethylacetate-heptane, isopropyl acetate-acetone, methyl-t-butylether-heptane, methyl-t-butyl ether-ethanol, ethanol-heptane,acetone-ethyl acetate, acetone-cyclohexane, toluene-heptane, morepreferably acetone.

Preferably, in step (ii) the basic compound of Cat is a compound capableof forming a salt with the acidic functionalities of the ARB and theNEPi. Examples include those mentioned above, such as calcium hydroxide,zinc hydroxide, calcium methoxide, calcium ethoxide, calcium acetate,calcium hydrogen carbonate, calcium formate, magnesium hydroxide,magnesium acetate, magnesium formate, magnesium hydrogen carbonate,sodium hydroxide, sodium carbonate, sodium hydrogen carbonate, sodiummethoxide, sodium ethoxide, sodium acetate, sodium formate, potassiumhydroxide, potassium carbonate, potassium hydrogen carbonate, potassiummethoxide, potassium ethoxide, potassium acetate, potassium formate,ammonium hydroxide, ammonium methoxide, ammonium ethoxide, and ammoniumcarbonate. Perchlorates may also be used. Amine bases or salt formingagents such as those mentioned above may also be used, in particularbenzathine, L-arginine, cholin, ethylene diamine, L-lysine orpiperazine. Typically an inorganic base is employed with Cat asspecified herein. More preferably, the basic compound is (Cat)OH,(Cat)₂CO₃, (Cat)HCO₃, still more preferably Cat(OH), such as NaOH. Thebasic compound is employed in an amount of at least 3 equivalentsrelative to either the ARB or the NEPi, preferably it is employed instoichiometric amount to obtain the dual-acting compound, in particularthe complex with three cations. The solvent used in step (ii) may be anysolvent or mixtures of solvents that allow dissolution of Cat(OH).Preferred solvents include water, methanol, ethanol, 2-propanol,acetone, ethyl acetate, isopropyl acetate, methyl-t-butylether,acetonitrile, toluene, and methylene chloride and mixtures of suchsolvents, more preferably water.

In step (iii) the solutions obtained in steps (i) and (ii) are combined.This can take place by adding the solution obtained in step (i) to thesolution obtained in step (ii) or vice versa, preferably, the solutionobtained in step (ii) to the solution obtained in step (i).

According to the first alternative, once combined and preferably mixed,the dual-acting compound, in particular the complex precipitates in step(iv). This mixing and precipitation is typically effected by stirringthe solutions for an appropriate amount of time such as 20 min to 6 h,preferably 30 min to 3 h, more preferably 2 h, at room temperature. Itis advantageous to add seeds of the dual acting compound. This methodfacilitates precipitation.

In step (iv) according to this first alternative, a co-solvent istypically added. The co-solvent employed is a solvent in which the ARBand the NEPi in the complexed form exhibit a lower solubility thatallows the compound to precipitate. Distillation, either continuous orstepwise, with replacement by this co-solvent results in a mixturepredominantly of the co-solvent.

Preferred solvents include ethanol, 2-propanol, acetone, ethyl acetate,isopropyl acetate, methyl-t-butylether, acetonitrile, toluene, andmethylene chloride and mixtures of such solvents, more preferablyisopropyl acetate. Preferably, a minimum amount of solvent is employedto facilitate precipitation. The solid is collected, e.g. by filtration,and is dried to obtain the dual-acting compound, in particular thecomplex in accordance with the present invention. The drying step can beperformed at room temperature or elevated temperature such as 30 to 60°C., preferably 30 to 40° C. Reduced pressure can be employed tofacilitate removal of the solvent, preferably, drying is effected atambient pressure or reduced pressure of e.g. 10 to 30 bar, such as 20bar.

According to a second alternative, once combined and preferably mixed,the dual-acting compound, in particular the complex the mixturepreferably forms a clear solution. This mixing is typically effected bystirring the solutions for an appropriate amount of time such as 20 minto 6 h, preferably 30 min to 3 h, more preferably 1 h, at roomtemperature. If necessary, the temperature may be raised so as to ensurea clear solution.

The obtained mixture is then further treated by solvent exchange toobtain the dual-acting compound, in particular the complex.

In step (iva) according to this second alternative, the solution ispreferably evaporated to dryness at elevated temperatures such as >roomtemperature to 50° C., more preferably 30 to 40° C.

Preferably, in step (va) the solvent or solvent mixture employed is asolvent in which the ARB and the NEPi in the complexed form exhibit alower solubility that allows the dual-acting compound, in particular thecomplex to precipitate. Preferred solvents include the ones mentionedabove for step (i), such as water, ethanol, 2-propanol, acetone ethylacetate, isopropyl acetate, methyl-t-butylether, acetonitrile, toluene,and methylene chloride and mixtures of such solvents, more preferablyisopropyl acetate. Preferably, a minimum amount of solvent or solventmixture is employed to facilitate precipitation.

In step (via) precipitation can take place at room temperature. It canbe effected by leaving the mixture standing or by agitating the mixture,preferably by agitating it. This is preferably effected by stirringand/or sonication. After precipitation, the solid is collected, e.g. byfiltration, and is dried to obtain the compound in accordance with thepresent invention. The drying step can be performed at room temperatureor elevated temperature such as 30 to 60° C., preferably roomtemperature. Reduced pressure can be employed to facilitate removal ofthe solvent, preferably, drying is effected at ambient pressure.

In a fourth aspect, this invention is directed to a method of treatingor preventing a disease or condition, such as hypertension, heartfailure (acute and chronic) congestive heart failure, left ventriculardysfunction and hypertrophic cardiomyopathy, diabetic cardiac myopathy,supraventricular and ventricular arrhythmias, atrial fibrillation,atrial flutter, detrimental vascular remodeling, myocardial infarctionand its sequelae, atherosclerosis, angina (unstable or stable), renalinsufficiency (diabetic and non-diabetic), heart failure, anginapectoris, diabetes, secondary aldosteronism, primary and secondarypulmonary hypertension, renal failure conditions, such as diabeticnephropathy, glomerulonephritis, scleroderma, glomerular sclerosis,proteinuria of primary renal disease, and also renal vascularhypertension, diabetic retinopathy, other vascular disorders, such asmigraine, peripheral vascular disease, Raynaud's disease, luminalhyperplasia, cognitive dysfunction (such as Alzheimer's), glaucoma andstroke comprising administering the afore-mentioned combination, linkedpro-drug or the dual-acting compound, in particular the complex to asubject in need of such treatment.

The combination, linked pro-drug or the dual-acting compound, inparticular the complex of the first embodiment may be administered aloneor in the form of a pharmaceutical composition according to the secondembodiment. Information regarding dosing, i.e., the therapeuticallyeffective amount, etc., is the same regardless of how the combination,linked pro-drug or he dual-acting compound, in particular the complex isadministered.

The combination, linked pro-drug or the dual-acting compound, inparticular the complex is beneficial over a combination of ARBs orneutral endopeptidase inhibitors alone or other ARB/NEPi combinationswith regard to use as first line therapy, ease of formulation and easeof manufacture.

Specific embodiments of the invention will now be demonstrated byreference to the following examples. It should be understood that theseexamples are disclosed solely by way of illustrating the invention andshould not be taken in any way to limit the scope of the presentinvention.

Example 1 Preparation of [valsartan((2R,4S)-5-biphenyl-4-yl-4-(3-carboxy-propionylamino)-2-methyl-pentanoicacid ethyl ester)]Na₃.2.5 H₂O

The dual-acting compound of valsartan and(2R,4S)-5-biphenyl-4-yl-4-(3-carboxy-propionylamino)-2-methyl-pentanoicacid ethyl ester is prepared by dissolving 0.42 g of(2R,4S)-5-biphenyl-4-yl-4-(3-carboxy-propionylamino)-2-methyl-pentanoicacid ethyl ester free acid (˜95% purity) and 0.41 g of valsartan freeacid in 40 ml acetone. Separately, 0.111 g of NaOH are dissolved in 7 mlH₂O. The two solutions are combined and stirred at room temperature for1 hour and a clear solution was obtained. The solution is evaporated at35° C. to yield a glassy solid. The glassy solid residue is then chargedwith 40 ml acetone and the resulting mixture is stirred and sonicateduntil precipitation occurred (˜5 minutes). The precipitate was filteredand the solid is dried at room temperature in open air for 2 days untila constant mass of the crystalline solid is obtained.

Characterization by various methods could confirm the presence of bothvalsartan and(2R,4S)-5-biphenyl-4-yl-4-(3-carboxy-propionylamino)-2-methyl-pentanoicacid ethyl ester and complex formation in contrast to a simple physicalmixture. Significant spectral peaks for the complex are observed e.g. inthe XRPD, IR, and Raman spectroscopy which are not present for thephysical mixture. See below for details on the characterization.

Example 2 Alternative Preparation of [valsartan((2R,4S)-5-biphenyl-4-yl-4-(3-carboxy-propionylamino)-2-methyl-pentanoicacid ethyl ester)]Na₃.2.5 H₂O

The dual acting compound of valsartan and(2R,4S)-5-biphenyl-4-yl-4-(3-carboxy-propionylamino)-2-methyl-pentanoicacid ethyl ester is prepared by dissolving 22.96 mmol of(2R,4S)-5-biphenyl-4-yl-4-(3-carboxy-propionylamino)-2-methyl-pentanoicacid ethyl ester free acid (˜95% purity) and valsartan (10.00 g; 22.96mmol) in acetone (300 mL). The suspension is stirred at room temperaturefor 15 min to obtain a clear solution. A solution of NaOH (2.76 g; 68.90mmol) in water (8 mL) water is then added to this solution over a periodof 10 min. Solids start to precipitate in 10 min. Alternatively,precipitation can be induced by seeding. The suspension is stirred at20-25° C. for 2 h. This suspension is concentrated at 15-30° C. underreduced pressure (180-250 mbar) to a batch volume of ˜150 mL. Isopropylacetate (150 mL) is then added to the batch and the suspension isconcentrated again at 15-30° C. under reduced pressure (180-250 mbar) toa batch volume of ˜150 mL. This operation (addition of 150 mL ofisopropyl acetate to the batch and concentration) is repeated onceagain. The suspension is stirred at 20-25° C. for 1 h. The solids arecollected by filtration under nitrogen over a Buchner funnel, washedwith isopropyl acetate (20 mL), and dried at 35° C. under reducedpressure (20 mbar) to afford the compound.

Characterization revealed the same product as in Example 1.

Example 3 Alternative Preparation of [valsartan((2R,4S)-5-biphenyl-4-yl-4-(3-carboxy-propionylamino)-2-methyl-pentanoicacid ethyl ester)]Na₃.2.5 H₂O Using Seeding

A reactor is charged with 2.00 kg (2,323 mmol) of AHU377 calcium saltand 20 L of isopropyl acetate. The suspension is stirred at 23±3° C.,and 4.56 L of 2 N HCl was added. The mixture is stirred at 23±3° C. for15 min to obtain a clear two-phase solution. The organic layer isseparated and washed with 3×4.00 L of water. The organic layer isconcentrated at 30-100 mbar and 22±5° C. to ˜3.5 L (3.47 kg) of AHU377free acid isopropyl acetate solution as a colorless solution.

To the above reactor containing ˜3.5 L (3.47 kg) of AHU377 free acidisopropyl acetate solution is added 1.984 kg (4,556 mmol) of Valsartanand 40 L of acetone. The reaction mixture is stirred at 23±3° C. toobtain a clear solution which is filtered into a reactor. To thereaction mixture is added a solution of 547.6 g (13,690 mmol) of NaOH in1.0 L of water at 23±3° C. (which was pre-cooled to 20±5° C. and in-linefiltered) over a period of 15-30 min while maintaining the internaltemperature at 20-28° C. (slightly exothermic). The flask is rinsed with190 mL of water and added into the reaction mixture. The reactionmixture is stirred at 23±3° C. for 15 min and a slurry of 4.0 g of[valsartan((2R,4S)-5-biphenyl-4-yl-4-(3-carboxy-propionylamino)-2-methyl-pentanoicacid ethyl ester)]Na₃.2.5 H₂O seeds in 50 mL of isopropyl acetate isadded. The mixture is stirred at 23±3° C. for 2 h to obtain asuspension. The suspension is heated to an internal temperature at 40±3°C. over a period of 20 min and 20 L of isopropyl acetate is added over aperiod of 20 min while maintaining the internal temperature at 40±3° C.The suspension is stirred at this temperature for an additional 30 min.The mixture is concentrated at an internal temperature at 35±5° C.(T_(j) 45±5° C.) under reduced pressure (200-350 mbar) to ˜35 L of awhite slurry (solvent collected: ˜25 L). Then 30 L of isopropyl acetateis added the mixture is concentrated at an internal temperature at 35±5°C. (T_(j) 45±5° C.) under reduced pressure (100-250 mbar) to ˜30 L of awhite slurry (solvent collected: ˜40 L). Again 40 L of isopropyl acetateis added and the mixture is concentrated at an internal temperature at35±5° C. (T_(j) 45±5° C.) under reduced pressure (100-200 mbar) to ˜30 Lof a white slurry (solvent collected: ˜30 L). The reaction mixture iscooled to 23±3° C. over ˜20 min and stirred at this temperature for anadditional 3 h. The solid is collected by filtration under nitrogen overa polypropylene pad on Buchner funnel. The solid is washed with 2×5 L ofisopropyl acetate and dried at 35° C. under reduced pressure (20 mbar)until isopropyl acetate content <0.5% to afford the above product as awhite solid.

Characterization revealed the same product as in Example 1.

X-Ray Powder Diffraction

Calculation of the interlattice plane intervals from the X-ray powderpattern taken with a Scintag XDS2000 powder diffractometer for the mostimportant lines for the sample give the following results:

d in [Å]: 21.2(s), 17.0(w), 7.1(s), 5.2(w), 4.7(w), 4.6(w), 4.2(w),3.5(w), 3.3(w)

The error margin for all interlattice plane intervals is ±0.1 Å. Theintensities of the peaks are indicated as follows: (w)=weak; (m)=medium;and (st)=strong.

Average values 2Θ in [°] are indicated (error limit of ±0.2) 4.5, 5.5,5.6, 9.9, 12.8, 15.7, 17.0, 17.1, 17.2, 18.3, 18.5, 19.8, 21.5, 21.7,23.2, 23.3, 24.9, 25.3, 27.4, 27.9, 28.0, 30.2.

Elemental Analysis

Elemental analysis gives the following measured values of the elementspresent in the sample. The findings of the elemental analysis, withinthe error limits, correspond to the overall formula of(C₄₈H₅₅N₆O₈Na₃).2.5H₂O

Found C: 60.05% H: 6.24% N: 8.80% Calculated* C: 60.18% H: 6.31% N:8.77%

Infrared Spectroscopy

The infrared absorption spectrum for the sample obtained usingAttenuated Total Reflection Fourier Transform Infrared (ATR-FTIR)spectrometer (Nicolet Magna-IR 560) shows the following significantbands, expressed in reciprocal wave numbers (cm⁻¹):

2956 (w), 1711 (st), 1637 (st), 1597 (st), 1488 (w), 1459 (m), 1401(st), 1357 (w), 1295 (m), 1266 (m), 1176 (w), 1085 (m), 1010 (w),942(w), 907 (w), 862 (w), 763 (st), 742 (m), 698 (m), 533 (st).

The error margin for all absorption bands of ATR-IR is ±2 cm⁻¹.

The intensities of the absorption bands are indicated as follows:(w)=weak; (m)=medium; and (st)=strong intensity.

Raman Spectroscopy

Raman spectrum of the sample measured by dispersive Raman spectrometerwith 785 nm laser excitation source (Kaiser Optical Systems, Inc.) showsthe following significant bands expressed in reciprocal wave numbers(cm⁻¹):

3061 (m), 2930 (m, broad), 1612 (st), 1523 (m), 1461 (w), 1427 (w), 1287(st), 1195 (w), 1108 (w), 11053 (w), 1041 (w), 1011 (w), 997 (m), 866(w), 850 (w), 822 (w), 808 (w), 735 (w), 715 (w), 669 (w), 643 (w), 631(w), 618 (w), 602 (w), 557 (w), 522 (w), 453 (w), 410 (w), 328 (w).

The error margin for all Raman bands is ±2 cm⁻¹.

The intensities of the absorption bands are indicated as follows:(w)=weak; (m)=medium; and (st)=strong intensity.

High Resolution CP-MAS ¹³C NMR Spectroscopy

The samples are investigated by high resolution CP-MAS (CrossPolarization Magic Angle Spinning) ¹³C NMR spectroscopy using aBruker-BioSpin AVANCE 500 NMR spectrometer equipped with a 300 Watt highpower ¹H, two 500 Watt high power X-amplifiers, necessary high powerpre-amplifiers, a “MAS” controller and a 4 mm BioSolids high resolutionBruker probe.

Each sample is packed in a 4 mm ZrO₂ rotor. Critical experimentalparameters are 3 msec ¹³C contact times, 12 KHz spinning speed at themagic angle, a “ramped” contact time, using a “SPINAL64” ¹H decouplingscheme, a recycle delay of 10 secs and 1024 scans at 293 deg K. Thechemical shifts are referenced with respect to an external Glycinecarbonyl at 176.04 ppm.

High resolution CP-MAS ¹³C NMR shows the following significant peaks(ppm):

179.0, 177.9 177.0, 176.7, 162.0, 141.0, 137.2, 129.6, 129.1, 126.7,125.3, 64.0, 61.5, 60.4, 50.2, 46.4, 40.6, 38.6, 33.5, 32.4, 29.8, 28.7,22.3, 20.2, 19.1, 17.8, 16.8, 13.1, 12.1, 11.1.

A physical mixture of individual Na salts of Valsartan and(2R,4S)-5-biphenyl-4-yl-4-(3-carboxy-propionylamino)-2-methyl-pentanoicacid ethyl ester revealed a simple inert mixture of the two salts.However, the sample of the complex prepared in Example 1 exhibiteddistinctly different spectral features in comparison to a 1:1 mixture ofthe sodium salts.

DSC and TGA

As measured by differential scanning calorimetry (DSC) using Q1000 (TAInstruments) instrument, the melting onset temperature and the peakmaximum temperature for the sample is observed at 139° C. and 145° C.,respectively.

As shown by DSC and thermogravimetric analysis (TGA), upon heating, thewater of hydration is released in two steps: the first step occurs below100° C. and the second step above 120° C.

Both DSC and TGA instruments are operated at a heating rate of 10 K/min.

Example 4 Preparation of Linked Pro-Drug of Scheme (1)

Linked pro-drug of valsartan calcium salt and(2R,4S)-5-biphenyl-4-yl-4-(3-carboxy-propionylamino)-2-methyl-pentanoicacid ethyl ester is prepared at room temperature by dissolving 114 mg ofthe calcium salt of valsartan and 86 mg of(2R,4S)-5-biphenyl-4-yl-4-(3-carboxy-propionylamino)-2-methyl-pentanoicacid ethyl ester free acid in 2 mL methanol, followed by methanolevaporation. The glassy solid residue is then charged with 3 mL ofacetonitrile and equilibrated by 10 min. sonication, followed by 20hours of magnetic stirring. Approximately 120 mg of white solids arecollected by filtration. Liquid chromatography (LC) and elementalanalysis indicate 1:1 ratio between(2R,4S)-5-biphenyl-4-yl-4-(3-carboxy-propionylamino)-2-methyl-pentanoicacid ethyl ester and valsartan. The sample is amorphous by X-ray powderdiffraction.

Preparation of Linked Pro-Drug of Scheme (2)

Linked pro-drug of valsartan calcium salt and(2R,4S)-5-biphenyl-4-yl-4-(3-carboxy-propionylamino)-2-methyl-pentanoicacid ethyl ester and Tris is prepared at room temperature by dissolving57 mg of the calcium salt of valsartan, 43 mg of(2R,4S)-5-biphenyl-4-yl-4-(3-carboxy-propionylamino)-2-methyl-pentanoicacid ethyl ester free acid, and 12.6 mg oftris(hydroxymethyl)aminomethane (Tris) in 2 mL methanol, followed bymethanol evaporation. The glassy solid residue is then charged with 3 mLof acetonitrile and equilibrated by 10 min. sonication, followed by 20hours of magnetic stirring. Approximately 83 mg of white solids arecollected by filtration. LC and elemental analysis indicate 1:1 ratiobetween(2R,4S)-5-biphenyl-4-yl-4-(3-carboxy-propionylamino)-2-methyl-pentanoicacid ethyl ester and valsartan. The sample is amorphous by X-ray powderdiffraction.

While the invention has been described above with reference to specificembodiments thereof, it is apparent that many changes, modifications,and variations can be made without departing from the inventive conceptdisclosed herein. Accordingly, it is intended to embrace all suchchanges, modifications and variations that fall within the spirit andbroad scope of the appended claims. All patent applications, patents,and other publications cited herein are incorporated by reference intheir entirety.

What is claimed is:
 1. An amorphous solid form of a compound consistingof anionic(S)—N-valeryl-N-{[2′-(1H-tetrazole-5-yl)-biphenyl-4-yl]-methyl}-valine,anionic(2R,4S)-5-biphenyl-4-yl-4-(3-carboxy-propionylamino)-2-methyl-pentanoicacid ethyl ester, and sodium cations in a 1:1:3 molar ratio.
 2. Apharmaceutical composition comprising the amorphous solid form accordingto claim 1 and at least one pharmaceutically acceptable excipient.
 3. Apharmaceutical composition suitable for oral administration comprisingsolid trisodium[3-((1S,3R)-1-biphenyl-4-ylmethyl-3-ethoxycarbonyl-1-butylcarbamoyl)propionate-(S)-3′-methyl-2′-(pentanoyl{2″-(tetrazol-5-ylate)biphenyl-4′-ylmethyl}amino)butyrate].4. A pharmaceutical composition suitable for oral administrationcomprising solid amorphous trisodium[3-((1S,3R)-1-biphenyl-4-ylmethyl-3-ethoxycarbonyl-1-butylcarbamoyl)propionate-(S)-3′-methyl-2′-(pentanoyl{2″-(tetrazol-5-ylate)biphenyl-4′-ylmethyl}amino)butyrate].5. A tablet prepared by a process comprising combining at least onepharmaceutically acceptable excipient with a solid amorphous form of: